The two-dimensional model of cell culture is an important method in the study of testicular development and spermatogenesis but can not effectively mimic and regulate the testicular microenvironment and the whole process of spermatogenesis due to the lack of relevant cell factors and the disruption of a three-dimensional spatial structure. In the past 20 years, the development and optimization of the in vitro model such as testis organotypic culture and in vivo model such as testis transplantation achieved a transformation from two- to three-dimension. The maintenance and optimization of the testicular niche structure could mimic the testicular microenvironment and cell types including Leydig, Sertoli and germ cells, which showed similar biological behaviors to those in vivo. Besides, the cell suspension or tissue fragment floats in the gas-liquid interface so that the development of somatic and germ cells is well maintained in vitro whilst the feedback linkage between grafted testis tissue and hypothalamus-pituitary of the host rebuilt in the in vitro model provides an endocrinological basis for spermatogenesis, which serves as an effective methodology to better understand the organogenesis and development of the testis as well as testicular function regulation, advancing the concept of treatment of male infertility. Al- though each of the methods may have its limitations, the progress in the processing, freezing, thawing, and transplantation of cells and tissues will surely promote their clinical application and present their value in translational medicine.
Cell Culture Techniques ; Germ Cells ; physiology ; Humans ; Infertility, Male ; therapy ; Male ; Spermatogenesis ; physiology ; Testis ; growth & development

Objective Investigated the treatment effect of ISCOM leukemia vaccine combination with 1-methyl tryptophan on tumor burden mice .Methods Saponin was added lipase protein (1 mg/mL) 7 ℃ for 12 h ,adding 80 μL lipid mixed solution and 5 mL saponin solution (1 mg/mL ) to prepare ISCOM leukemia vaccine .C57BL /6 mice were randomly divided into model group , ISCOM leukemia vaccine group ,1-methyl tryptophan group and combination group ,Mice were injected FBL-3 cell to built leukemia tumor-burdened mice model .After treatment for 4 weeks ,tumors weight ,NK and Mφ and CTL cell killing activity ,serum levels of IL-10 ,IL-12 were detected .Results Tumor weight in combination group was less than 1-methyl tryptophan and ISCOM leukemia group [(0 .64 ± 0 .26)g vs .(2 .49 ± 0 .91)g ,P< 0 .01 ,(0 .64 ± 0 .26)g vs .(1 .28 ± 0 .73)g ,P< 0 .05] ;NK cell killing activity in com-bination group was higher than 1-methyl tryptophan group[(38 .41 ± 8 .27)% vs .(67 .22 ± 12 .74)% ,all P< 0 .01)] ;M φ activity in combination group was significantly higher than 1-methyl tryptophan group[(55 .69 ± 13 .69)% vs .(69 .47 ± 14 .79)% ,P< 0 .01] ;CTL activity in combination group was significantly higher than 1-methyl tryptophan group and ISCOM leukemia group[(43 .77 ± 8 .89)% vs .(69 .68 ± 11 .44)% ,P< 0 .01 ,(58 .87 ± 9 .45)% vs .(69 .68 ± 11 .44)% ,P < 0 .05] ;IL-10 in combination group were significantly lower than 1 - methyl tryptophan group and ISCOM leukemia group [(76 .2 ± 6 .82)pg /L vs .(98 .3 ± 13 .4)pg/L ,P<0 .01 ,(76 .2 ± 6 .82)pg/L vs .(202 .3 ± 44 .5)pg/L ,P < 0 .01] ;IL-12 in combination group were significantly higher than 1-methyl tryptophan group and ISCOM leukemia group[(381 .2 ± 47 .3)pg/L vs .(332 .1 ± 30 .2)pg/L ,P < 0 .05 ,(381 .2 ± 47 .3)pg /L vs . (291 .2 ± 17 .3)pg/L ,P< 0 .01] .Conclusion Combination with 1-methyl tryptophan and ISCOM leukemia vaccine has a well anti-tumor effect ,its mechanism may be through mediated and the expression of IL-12 and IL-10 .

Objective To explore the causes and treatment of repeated vulvovaginitis in girlhood in order to improve its prevention and treatment.Methods Fifty-one girls with repeated vulvovaginitis(age≤10 years)admitted to The Second Affiliated Hospital of Sun Yat-sen University from Jan.1990 to Nov.2004 were reviewed retrospectively.Results We found 28 girls(55%)suffering from non-specific vulvovaginitis and 14 ones(27%)suffering from posterior recto-vaginal fistula with in 51 patients.Five girls(10%)were smitten with vulval ulcer and 3 ones(6%)had been were found with vaginal foreign bodies.One girl(2%) was smitten with adhesion of labia minora.The vaginal discharges taken from 21 girls were cultured. Seventeen cases found bacteria.The positive rate of bacteria culture in the 21 cases reached 81%,in which, E.coli accounted for 5 cases(24%),staphylococcus and streptococcus accounted for 3 cases(14%) respectively.Patients suffered from non-specific vulvovaginitis and vulval ulcer accepted external lotion, antibiotic ointment or combining with antibiotics.Patients suffered from posterior recto-vaginal fistula accepted fistulectomy.Three girls who found vaginal foreign bodies took out of foreign bodies by hysteroscopo.Fifty-one girls all were cured after appropriate therapy.Conclusions Vulvovaginitis is the most common gynecologic diagnosis in girlhood.The principal cause of repeated invasion is non-specific vulvovaginitis and the secondly one is posterior recto-vaginal fistula.It need overhaul during the diagnosis.It is very availability to use hysteroscopy and do bacteria culture+antibiotic sensitivity test for repeated pediatric vulvovaginitis.

HCV RNA positive serum was first selected by RT-PCR test kit from several anti-HCV positive sera obtained from Xi'an.HCV RNA extrac ted from the elected sera was converted to cDNA by reverse transcription with ra ndom primer.Half-nested PCR was performed.The amplified product was 852 bp.The purified PCR product was digested by restriction endonucleases and then ligated to epressio vector pET-22b\++.Its nucleotide sequence was determined by dideoxy chain termination method.A comparison of the sequence with several isolates rep orted previously showed that the sequence belonged to HCV type Ⅱ.

With dwindling number of new antibiotics and inappropriate use of antibiotics, the emergence and spread of antibiotics resistance occurs commonly in healthcare institutions worldwide. In China, antibiotics are commonly overprescribed and misused. This study is to assess the effect of the nationwide special campaign on antibiotic stewardship program (ASP) at specialized hospitals in China by investigating prescription information from 2011 to 2012. Data on the hospital consumption and prescription of systemic antibiotics were obtained from four specialized hospitals, including maternity, children's, stomatological and cancer hospitals. Systematic random sampling was used to select outpatient prescriptions and inpatient cases. A total of 105 specialized hospitals in 2011 and 121 specialized hospitals in 2012 were analysed. The defined daily doses (DDDs) per 100 inpatient days, the percentage of antibiotic use in outpatient prescriptions, and the percentage of antibiotic use in inpatient cases were used as measurements of antibiotic use. The overall antibiotic use density in the selected hospitals decreased between 2011 and 2012 from 39.37 to 26.54 DDD/100 inpatient days (P<0.001). The percentage of antibiotic use in outpatient prescriptions (range: 24.12%-18.71%, P=0.109) and inpatient cases (64.85%-60.10%, P=0.006) also decreased within the two years. Significant changes were observed among regions and different hospitals within the two years. And antibiotic consumption was correlated with the type and size of specialized hospital in 2012, but not with the regions. This analysis of antibiotic consumption of specialized hospitals allows relevant comparisons for benchmarking and shows that national ASP has improved antibiotic rational use in China. The data will assist policymakers in formulating effective strategies to decrease antibiotic overuse and identify areas that require further work.

Aim To observe the antifibrotic effect of Radis Astragali and to explore the mechanism. Methods The fibrosis of animal model was induced with CCl4,and the model animals were treated with Radis Astragali in treatment group and saline in control group respectively. Results The serum hyaluronic acid (HA), fibrosis score and ICAM_1_positive hepatocytes all the decreased in the treatment group as compared to those in the control group.Conclusion Radis Astragali has satisfactory effect on experimental fibrosis. The mechanism may be correlated with its affection on ICAM_1 in liver tissue.

Objective To investigate the point mutations within the adenosine triphosphate-binding region of BCR/ABL in patients with chronic myeloid leukemia who develop imatinib resistance.Methods We collected a total of 17 bone marrow samples obtained from 11 patients who showed hematology resistance (n = 7)or cytogenetic refractoriness(n = 4).A long semi-nest PCR method was used to amplify the ABL kinase domain of the BCR/ABL allele.After two rounds of PCR reactions,we got a fragment of 863 bases The PCR products were purified and followed by sequencing.Results In total,we find three point mutations presented in all patients tested G250E,E255K and T315I.The mutation rate of hematology resistance is 4/ 7,and 95% confidence interval was 8%-90%,while mutation rate of cytogenetic refractoriness 1/4,95% confidence interval 1%-81%.For those patients whose samples were available,no single mutation were determined before imatinib resistance emerged.Conclusions There is high frequency of point mutations clustered within the adenosine triphosphate-binding region of BCR/ABL in patients with chronic myeloid leukemia.It's good for patients to switch to another therapeutic strategy when the mutations are detected.

Cathepsin B from Helicoverpa armigera (HCB) belongs to the group of cysteine proteinases. HCB is proved being involved in the degradation of yolk proteins during embryonic development,which is an acidic preferring enzyme and is resistant to SDS. The expression of the proenzyme may offer a model for investigating the activation of the enzyme. The HCB gene was constructed into pPIC9K and expressed in Pichia pastoris KM71 strain . After induction by methanol, HCB was expressed and secreted into the medium. The molecular weight of the recombinant procathepsin B was determined as about 38 kDa. The expressed product was confirmed to be HCB by immunoblotting assay using specific rabbit anti-HCB polyclonal antibody. The activity of the product was assayed by in situ hydrolysis (gelatin-SDS-PAGE). These results showed that HCB with proteolytic activity was expressed in P. pastoris KM71. This proenzyme can be used for further research on the activation of the proenzyme or industrial production.

Objective To investigate the effect of arsenic trioxide(As_2 O_3)nanoparticles on rabbit vascular smooth muscle cells in vitro in comparison with normal form As_2 03.Methods The rabbit vascular smooth muscle cells were cultured in vitro.Nano and normal forms of As_2O_3 with drug concentrations of 3?mol/L were added into the cells.Cell proliferation curve was drawn according to the light absorption values of MTT test.Flow cytometry was applied to observe the apoptosis.DNA was extracted and underwent electrophoresis.Results Cell proliferation treated with the 3?mol/L concentration of As_2O_3 was inhibited. Cell growth was inhibited markedly with increased treatment time,and the inhibition effect of nano drug form seemed stronger than that of normal form.MTT light absorption values of cells treated at 24,48 and 72 h showed statistically significant difference(H=10.934,15.039,15.539,P

OBJECTIVETo study the effect of nitric oxide (NO) in vestibular compensation after unilateral vestibular deafferentation.

METHODSEighteen animals were divided into two groups, 6 of group a as control, 12 of group b received gentamicin intratympanic injection in the left ear. Half of the animals were killed respectively after 5 days and 10 days. Vestibular endorgan and brainstem tissue sections were subjected to NADPH-d reactive test of NOS for histochemical examination.

RESULTSIn group a, NOS-like reactivity in both sides of vestibular endorgan and nucli. In group b during 5 days, NOS-like reactivity in right side of vestibular endorgan and nucli, those of the left side were negative. During 10 days, NOS-like reactivity only in the right side of vestibular endorgan.

CONCLUSIONSChanges of NOS expression in the contralateral vestibular nucli might have played a role in vestibular compensation.

Adaptation, Physiological ; Animals ; Brain Stem ; metabolism ; physiopathology ; Evoked Potentials, Auditory, Brain Stem ; Female ; Gentamicins ; Guinea Pigs ; Male ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Vestibular Nerve ; metabolism ; Vestibular Nuclei ; metabolism ; physiopathology ; Vestibule, Labyrinth ; injuries ; metabolism ; physiopathology